6.1.3 Manipulating Genomes

Question 1

why is polymerase chain reaction (PCR) done?

Answer 1

to amplify fragments of DNA

Question 2

how does PCR work?

Answer 2

1. a reaction mixture set up that contains DNA sample, free nucleotides, primers and DNA polymerase
2. primers are short pieces of DNA that are complimentary to bases at the start of fragment
3. DNA polymerase creates new DNA strands
4. DNA mixture heated to 95°C, to break H bonds between bases
5. **mixture then cooled to 50-65 °C **so primers can bind
6. reaction mixture heated to 72°C, so DNA polymerase can work
7. DNA polymerase lines up free DNA nucleotides alongside each template strand, they pair by complimentary base pairing
8. two new copies of fragment of DNA are formed and one cycle of PCR is complete
9. each PCR cycle doubles amount of DNA

Question 3

what does sequencing DNA mean and why’s it done?

Answer 3

to determine the order of bases in a section of DNA

Question 4

what’s the og way to sequence DNA?

Answer 4

Sanger technique

Question 5

how do you carry out the Sanger technique of sequencing?

Answer 5

1. extract DNA you wanna sequence and cut it into fragments of various lengths
2. amplify (make many copies) the DNA by PCT
3. add this DNA to 4 different solutions (A,G,C,T)
4. these 4 diff solutions contain DNA nucleotides and DNA polymerase and primers and terminator base (A, C, G, T)
5. If you understand later let me know 🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣🤣

Question 6

what are new, faster ways for DNA sequencing?

Answer 6

• high throughput sequencing
• next generation sequencing
• massive parallel sequencinv